Model Call Girl in Bikash Puri Delhi reach out to us at 🔝9953056974🔝
Partial thesis defence presentation
1. Structural Biology of Cytochrome P450 monooxygenases in Arabidopsis thaliana phenylpropanoid pathway Sanjeewa Rupasinghe Mary Schuler lab Department of Cell and Developmental Biology spring 2007
2. Cytochrome P450 monooxygenases Use molecular O 2 and NADPH to catalyze monooxygenation reactions R-H + O 2 + NADPH R-OH + H 2 O + NADP Catalyze key steps in metabolic pathways from archaea to mammals Responsible for drug metabolism in humans Responsible for insecticide metabolism in insects wavelength (nm)
6. phenylpropanoid P450s OH OH O OH O CYP73A5 OH O OH OH OH OH O OH OH CYP84A1 O O OH OH OH OH O O OH OH OH OH OH CYP75B1 O O OH COOH OH OH O O OH COOH OH OH OH CYP98A3
7.
8. Structural biology of P450s in A. thaliana phenylpropanoid pathway Homology modeling and substrate docking Mutagenesis and protein engineering X-ray crystallography
14. Kyoto encyclopedia of Genes and Genomes ( www.kegg.com ) 11000 + biological compounds Other databases 3000+ compounds List of most possible substrates Arabidopsis thaliana P450s 272 + genes Functions known for only 40 http://arabidopsis-p450.biotec.uiuc.edu Homology modeling High throughput docking
15.
16.
17.
18. Protein engineering in CYP98A3 CYP98A3 CYP73A5 O O OH OH O OH OH O OH OH O OH OH O O 292 302 381 382 473 479 301 309 368 375 484 489 1 CYP98A3 CYP73A5
19.
20. X-ray crystallographic studies of phenylpropanoid P450s CYP98A3 CYP98A3wt MSWFLIAVATIAAVVSYKLI QRLRYKFPPG PSPKPIVGNLYDIKP VRFRCYYE CYP98A3D1 MA------------------QRLRYKFPPGPSPKPIVGNLYDIKPVRFRCYYE CYP98A3D2 MA------------------QR R R R KFPPGPSPKPIVGNLYDIKPVRFRCYYE promoter/ E. coli strain
23. Structural Biology of P450s in A. thaliana phenylpropanoid pathway Homology modeling Mutagenesis and Protein engineering X-ray crystalography NMR
24. Solid state NMR studies of Phenylpropanoid P450s Collaboration with Chad Rienstra lab (UIUC) Labeling P450s with C 13 and N 15 (Rupasinghe et al submitted to chem biochem journal) Promoter / Chaperone co-expression System P450 yield (mg/L of culture) Minimum Media Terrific Broth tac promoter ND 3 Bacteriopage T7 promoter ND 3 tac promoter / araB groES-groEL 4 8 tac promoter / Pzt1 groES-groEL 2 6 Spectra acquired by Heather Fererics , Chad Rienstra lab